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  • Self-immolative polycations as gene delivery vectors and prodrugs targeting polyamine metabolism in cancer.

Self-immolative polycations as gene delivery vectors and prodrugs targeting polyamine metabolism in cancer.

Molecular pharmaceutics (2014-08-26)
Yu Zhu, Jing Li, Shrey Kanvinde, Zhiyi Lin, Stuart Hazeldine, Rakesh K Singh, David Oupický
ABSTRACT

Polycations are explored as carriers to deliver therapeutic nucleic acids. Polycations are conventionally pharmacological inert with the sole function of delivering therapeutic cargo. This study reports synthesis of a self-immolative polycation (DSS-BEN) based on a polyamine analogue drug N(1),N(11)-bisethylnorspermine (BENSpm). The polycation was designed to function dually as a gene delivery carrier and a prodrug targeting dysregulated polyamine metabolism in cancer. Using a combination of NMR and HPLC, we confirm that the self-immolative polycation undergoes intracellular degradation into the parent drug BENSpm. The released BENSpm depletes cellular levels of spermidine and spermine and upregulates polyamine catabolic enzymes spermine/spermidine N(1)-acetyltransferase (SSAT) and spermine oxidase (SMO). The synthesized polycations form polyplexes with DNA and facilitate efficient transfection. Taking advantage of the ability of BENSpm to sensitize cancer cells to TNFα-induced apoptosis, we show that DSS-BEN enhances the cell killing activity of TNFα gene therapy. The reported findings validate DSS-BEN as a dual-function delivery system that can deliver a therapeutic gene and improve the outcome of gene therapy as a result of the intracellular degradation of DSS-BEN to BENSpm and the subsequent beneficial effect of BENSpm on dysregulated polyamine metabolism in cancer.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Spermidine, BioUltra, for molecular biology, ≥99.5% (GC)
Sigma-Aldrich
5-Sulfosalicylic acid dihydrate, purum p.a., ≥98.0% (T)
Sigma-Aldrich
Hexylene glycol, puriss., ≥99.0% (GC)
Sigma-Aldrich
Spermine, ≥99.0% (GC)
Sigma-Aldrich
Hexylene glycol, BioUltra, ≥99.0% (GC)
Sigma-Aldrich
Dichloromethane, suitable for HPLC, ≥99.9%, contains 40-150 ppm amylene as stabilizer
Sigma-Aldrich
Hexylene glycol, 99%
Sigma-Aldrich
L-Glutamine, BioUltra, ≥99.5% (NT)
Supelco
Dichloromethane, Selectophore, ≥99.5%
Sigma-Aldrich
1,6-Hexanediol, 97%
Sigma-Aldrich
5-Sulfosalicylic acid dihydrate, ACS reagent, ≥99%
Sigma-Aldrich
Dichloromethane, anhydrous, ≥99.8%, contains 40-150 ppm amylene as stabilizer
Sigma-Aldrich
Ethylenediaminetetraacetic acid, 99.995% trace metals basis
Sigma-Aldrich
Ethylenediaminetetraacetic acid, ≥98.0% (KT)
Sigma-Aldrich
Ethylenediaminetetraacetic acid, BioUltra, ≥99.0% (KT)
Sigma-Aldrich
Ethylenediaminetetraacetic acid disodium salt solution, BioUltra, for molecular biology, pH 8.0, ~0.5 M in H2O
Sigma-Aldrich
L-Glutamine
Supelco
L-Glutamine, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Supelco
L-Glutamine, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Dichloromethane solution, contains 10 % (v/v) methanol
Sigma-Aldrich
HEPES buffer solution, 1 M in H2O
Supelco
Dichloromethane, analytical standard
Sigma-Aldrich
Spermidine, ≥99% (GC)
Sigma-Aldrich
Spermine, suitable for cell culture, BioReagent
Sigma-Aldrich
Spermine, ≥97%
Sigma-Aldrich
5-Sulfosalicylic acid dihydrate, suitable for electrophoresis, ≥99%
Sigma-Aldrich
Ethylenediaminetetraacetic acid, anhydrous, crystalline, BioReagent, suitable for cell culture
Sigma-Aldrich
Ethylenediaminetetraacetic acid, purified grade, ≥98.5%, powder
Sigma-Aldrich
Ethylenediaminetetraacetic acid, ACS reagent, 99.4-100.6%, powder
Sigma-Aldrich
L-Glutathione reduced, BioXtra, ≥98.0%